In vitro translation is an important tool for research in molecular biology, diagnostic assays, and biological discovery. Commercially available cell free lysate kits work by supplying a cell free lysate that contains most if not all of the cellular components necessary to translate mRNA to which the user adds DNA or mRNA encoding a protein of interest. These in vitro translation kits have enabled biological discovery for important ramifications in health care, drug design, and biological sensing. Under conditions suitable for translation to occur, the protein of interest is synthesized using translation machinery provided by the lysate. However, one serious limitation of currently available methods of in vitro translation is the amount of protein that is produced. Many kits and/or methods suffer from low and unreliable protein production. Because mRNA can be easily degraded by numerous mechanisms, mRNA lifetime is viewed as one limiting factor of current in vitro translation methods. Current strategies for improving protein production typically focus on increasing mRNA stability and resistance to degradation. However, translation is a complex process, involving not only mRNA but also multiple translation factors, all of which must be coordinated. In particular, initiation, a complex first step of translation, requires numerous initiation factors to be brought in close proximity of the mRNA, the ribosome, and one another. As a result the process in vitro is very inefficient and produces little protein.
As shown in FIG. 2, the biological process of translation, that is, synthesis of a specific protein encoded in a particular mRNA is a complicated process involving hundreds of different components.5 The different components interact with each other and the mRNA in a coordinated fashion to synthesize the protein encoded by the mRNA.
Translation can be conducted in the absence of cells (in vitro) and is a central biological tool that is used in many areas of bioengineering and biological research. In vitro translation is used to produce specific proteins of interest from nucleic acid sequences encoding them, in the engineering of proteins for pharmaceutical, diagnostic, and research tool use, as well as in the search and discovery of proteins. To reproduce this complicated biological process in the laboratory setting, the many biological components shown in FIG. 2 must be provided to the reaction. Such translation components are provided in the form of kits. However, even with the use of kits, in vitro translation is highly inefficient and typically produces very small quantities of protein product.